Distribution of parasite cysteine proteinases in lesions of mice infected with Leishmania mexicana amastigotes

Ilg, Thomas; Fuchs, Manuela; Gnau, Volker; Wolfram, Markus; Harbecke, Dorothee; Overath, Peter

doi:10.1016/0166-6851(94)00126-x

Cited by 38 publications

(26 citation statements)

References 33 publications

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“…The first was a family of CPs (24 to 27 kDa) that is abundantly expressed in the lysosomes of amastigotes (3.5% of the cellular protein corresponding to 1.2 ϫ 10 6 molecules/cell) but not in promastigotes. These enzymes are encoded by the lmcpb gene family (24,25,38,39). The second was a zinc metalloproteinase, a glycoprotein of 63 kDa (GP63) carrying a glycosylphosphatidylinositol (GPI) membrane anchor, which is the major surface protein of promastigotes (0.5 to 1% of the cellular protein corresponding to about 5 ϫ 10 5 molecules/cell); this protein is present in promastigotes of all Leishmania species investigated (53).…”

Section: Resultsmentioning

confidence: 99%

“…Native antigens. Cysteine proteinases (CP) were purified from lesion-derived amastigotes or from the supernatant of lesion homogenates (24,25). The isolation of GP63-related proteins (GP63) from amastigotes and a truncated form of MBAP from the culture supernatant of promastigotes has been described before (24,68).…”

Section: -Mrgshhhhhhgs (Vector)-ykvel-------aiiv-cooh (Rmbap)mentioning

confidence: 99%

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Subunit Vaccination of Mice against New World Cutaneous Leishmaniasis: Comparison of Three Proteins Expressed in Amastigotes and Six Adjuvants

et al. 2000

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A mixture of well-defined recombinant antigens together with an adjuvant that preferentially stimulates specific gamma interferon (IFN-␥)-secreting helper type 1 CD4 ؉ T cells (Th1 cells) presents a rational option for a vaccine against leishmaniasis. The potential of this approach was investigated in murine infections with Leishmania mexicana, which are characterized by the absence of a parasite-specific Th1 response and uncontrolled parasite proliferation. A mixture of three antigens (glycoprotein 63, cysteine proteinases, and a membrane-bound acid phosphatase), which are all expressed in amastigotes, the mammalian stage of the parasite, were used for the immunization of C57BL/6 mice in combination with six adjuvants (interleukin 12 [IL-12], Detox, 4-monophosphoryl lipid A, QS-21, Mycobacterium bovis BCG, and Corynebacterium parvum). All six vaccine formulations containing the mixture of recombinant antigens were protective against challenge infections with promastigotes, the insect stage of the parasite, in that mice controlled and healed infections but developed transient and, in certain cases, accentuated disease. The most effective adjuvants were IL-12 followed by Detox. Further studies using these two adjuvants showed that a similar protective effect was observed with a mixture of the corresponding native proteins, and mice which had controlled the infection showed a preponderance of IFN-␥-secreting CD4 ؉ T cells in the lymph nodes draining the lesion. Using the recombinant proteins individually, it is shown that the relatively abundant cysteine proteinases and glycoprotein 63, but not the acid phosphatase, are able to elicit a protective response. The results are discussed in comparison to previous studies with subunit vaccines and with respect to cell biological aspects of antigen presentation in Leishmania-infected macrophages.The solid immunity observed following recovery from cutaneous leishmaniasis in humans has stimulated numerous attempts for the development of a prophylactic vaccination against this widespread (sub)tropical disease (12,16,19,35,36). Leishmania major, the etiological agent of Old World cutaneous leishmaniasis, has been the most popular species both in studies of murine infections and in human trials. In the Middle East, the deliberate infection with L. major was a common and effective practice for immunization against subsequent infections, but a fraction of the vaccinated persons produced lesions that required medical treatment. A vaccine based on killed promastigotes, the insect stage of the parasite, and Mycobacterium bovis BCG was recently shown to be ineffective in a controlled trial with several thousand volunteers in Iran (37). Mice have been used in a range of vaccination protocols against infection by L. major. Representative examples include: attenuated but live parasites (23,29,34,48,59); subunit vaccines delivered by live carriers such as BCG expressing the surface proteinase GP63 of L. major (11); vaccinia virus expressing the glycoprotein GP46/M-2 (20, 30); GP63 expr...

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Section: Resultsmentioning

confidence: 99%

Section: -Mrgshhhhhhgs (Vector)-ykvel-------aiiv-cooh (Rmbap)mentioning

confidence: 99%

Subunit Vaccination of Mice against New World Cutaneous Leishmaniasis: Comparison of Three Proteins Expressed in Amastigotes and Six Adjuvants

et al. 2000

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“…The CPB genes of L. mexicana are stage-regulated cathepsin L-like enzymes, with 2 of the 19 genes in the array being expressed in infective metacyclic promastigotes, while the other 17 are predominantly expressed in amastigotes (7); thus, expression occurs exclusively in parasite stages that interact with the mammalian host. Although these CPB enzymes are found in the megasomes (extended, large lysosomes), they have been found in the parasitophorous vacuole and extracellularly in lesions (14), giving the enzymes ample opportunity to interact with host proteins.…”

Section: Discussionmentioning

confidence: 99%

“…L. mexicana possesses three types of CPs of the papain superfamily: CPA and CPB have homology to mammalian cathepsin L; CPC has homology to mammalian cathepsin B, but has substrate specificity closer to cathepsin L. CPB is especially abundant, and the 19 CPB genes are stage regulated, with most having the greatest expression in amastigotes (the parasite stage that occurs in the mammalian host). These enzymes have been localized to the megasomes (large extended lysosomes found in L. mexicana, but not L. major), but are also present in the parasitophorous vacuole, and in abundance in the extracellular tissue of murine leishmaniasis lesions (14). Although L. major contains a homologous array of CPB genes, CP activity is considerably lower than in L. mexicana (15), which shows that there are major differences in enzyme levels and/or specificities between these two Leishmania species.…”

Section: Eishmania Mexicana Like Other New World Leishmaniamentioning

confidence: 99%

Cysteine Protease B of Leishmania mexicana Inhibits Host Th1 Responses and Protective Immunity

Buxbaum

Denise

Coombs

et al. 2003

The Journal of Immunology

108

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C3H mice infected with Leishmania mexicana fail to develop a protective Th1 response, and are unable to cure. In this study, we show that L. mexicana cysteine proteases suppress the antileishmanial immune response. Previous studies demonstrated that deletion of the entire multicopy cysteine protease B (CPB) gene array in L. mexicana is associated with decreased parasite virulence, potentially attributable to factors related to parasite fitness rather than to direct effects on the host immune response. We now show that C3H mice infected with the L. mexicana deletion mutant (Δcpb) initially develop lesions that grow at rates comparable to those of wild-type L. mexicana-infected mice. However, in contrast to controls, Δcpb-induced lesions heal with an accompanying Th1 immune response. Lesion resolution was Th1 dependent, as Δcpb-infected IL-12p40−/− and STAT4−/− mice developed high parasite burdens and progressive disease. Moreover, when L. major was transfected with a cosmid expressing multiple L. mexicana CPB genes, this parasite induced a significantly lower IFN-γ response compared with wild-type L. major. These data indicate that cysteine proteases of L. mexicana are critical in suppressing protective immune responses and that inhibition of CPB may prove to be a valuable immunomodulatory strategy for chronic forms of leishmaniasis.

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“…At least 50% of the cysteine peptidase activity is localized within the lysosomes of the parasite, so is unlikely to pay a direct role in modulating macrophage signaling (69). However, it is also clear that a significant number of cysteine peptidases are released following amastigote death and as a result of their unusual intracellular trafficking pathway (17,69). As described below, these released proteases can modulate macrophage activity by acting directly on the host cell surface or following entry into the macrophage endoplasmic reticulum from the phagosome.…”

Section: Parasite Surface and Secreted Moleculesmentioning

confidence: 99%

Subversion Mechanisms by WhichLeishmaniaParasites Can Escape the Host Immune Response: a Signaling Point of View

2005

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The obligate intracellular parasite Leishmania must survive the antimicrobial activities of its host cell, the macrophage, and prevent activation of an effective immune response. In order to do this, it has developed numerous highly successful strategies for manipulating activities, including antigen presentation, nitric oxide and oxygen radical generation, and cytokine production. This is generally the result of interactions between Leishmania cell surface molecules, particularly gp63 and LPG, and less well identified macrophage surface receptors, causing the distortion of specific intracellular signaling cascades. We describe some of the signaling pathways and intermediates that are repressed in infected cells, including JAK/STAT, Ca2+-dependent protein kinase C (PKC) isoforms, and mitogen-activated protein kinases (especially ERK1/2), and proteasome-mediated transcription factor degradation. We also discuss protein tyrosine phosphatases (particularly SHP-1), intracellular Ca2+, Ca2+-independent PKC, ceramide, and the suppressors of cytokine signaling family of repressors, which are all reported to be activated following infection, and the role of parasite-secreted cysteine proteases

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Distribution of parasite cysteine proteinases in lesions of mice infected with Leishmania mexicana amastigotes

Cited by 38 publications

References 33 publications

Subunit Vaccination of Mice against New World Cutaneous Leishmaniasis: Comparison of Three Proteins Expressed in Amastigotes and Six Adjuvants

Subunit Vaccination of Mice against New World Cutaneous Leishmaniasis: Comparison of Three Proteins Expressed in Amastigotes and Six Adjuvants

Cysteine Protease B of Leishmania mexicana Inhibits Host Th1 Responses and Protective Immunity

Subversion Mechanisms by WhichLeishmaniaParasites Can Escape the Host Immune Response: a Signaling Point of View

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